Structural and functional characterization of ATP synthase c-subunit leak channel and its role in AD pathogenesis

ATP合酶c亚基泄漏通道的结构和功能特征及其在AD发病机制中的作用

基本信息

批准号：
10214094
负责人：
Nelli Mnatsakanyan
金额：
$ 15.81万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-15 至 2022-01-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10214094
关键词：
Affinity Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Alzheimer&apos s disease pathology Amyloid beta-Protein Architecture Binding Biology Brain Calcium Cell Death Cell Death Induction Cell Survival Cells Cleaved cell Complex Cryo-electron tomography Cryoelectron Microscopy Detergents Development Energy Metabolism Environment Event Functional disorder Glutamates Goals Hippocampus (Brain)Human Immobilization In Situ Individual Inner mitochondrial membrane Interferometry Kinetics Lipids Liposomes Literature Measures Membrane Mitochondria Mitochondrial Proton-Translocating ATPases Modeling Molecular Molecular Conformation Mus N-terminal Neurodegenerative Disorders Neurons Oligomycins Outer Mitochondrial Membrane Pathogenesis Pathologic Pathway interactions Permeability Pharmacology Play Prevention strategy Property Proteins Reporting Research Resolution Role Rupture Signal Transduction Structure Synapses System Testing Toxic effect Transgenic Mice cyclophilin D cytochrome c driving force inhibitor/antagonist mitochondrial dysfunction mitochondrial membrane mitochondrial permeability transition pore monomer mouse model nanodisk novel novel therapeutics overexpression particle patch clamp prevent reconstitution sensor time use

项目摘要

Abstract Mitochondrial and synaptic dysfunction are early pathological features and a driving force of Alzheimer's disease (AD) pathology. Aβ is found to accumulate abnormally in the brains of AD individuals and in an AD mouse models leading to mitochondrial Ca2+ overload and activation of mitochondrial permeability transition pore (mPTP). Prolonged opening of mPTP triggers outer mitochondrial membrane rupture, release of cytochrome c and activation of downstream cell death pathways. The mPTP has been at the center of extensive scientific research for the last several decades but it still remains as one of the most mysterious phenomena in biology today due to its controversial molecular composition and the lack of structural information of its pore. We have recently demonstrated the novel role of ATP synthase c-subunit ring in forming the channel of mPTP. Nevertheless, the gating mechanism of the ATP synthase c-subunit leak channel and conformational changes initiating its opening are yet to be discovered. In this proposal, we will use single-particle cryo-electron microscopy (cryo-EM) to identify the high- resolution structure and the open conformation of ATP synthase leak channel in the presence of channel modulators. We will also perform in situ structural analysis of ATP synthase in its functional environment within the Aβ-exposed primary hippocampal neurons and in mitochondria isolated from the mouse models of AD by using cryo-electron tomography (cryo-ET). In this project we will also investigate the direct role of ATP synthase leak channel as a novel cell death pathway in AD pathogenesis; we will test whether the pharmacological inhibition of this channel will rescue neurons from Aβ-induced cell death. Successful completion of this proposal will reveal the molecular mechanism(s) of mitochondrial permeability transition, the atomic structure of ATP synthase leak channel, and will aid in the development of new treatments for AD, targeting ATP synthase.

抽象的线粒体和突触功能障碍是早期病理特征，也是疾病发生的驱动力。阿尔茨海默病 (AD) 病理学发现 Aβ 在 AD 患者的大脑中异常积累。个体和 AD 小鼠模型中导致线粒体 Ca2+ 过载和激活线粒体通透性转换孔 (mPTP) 的延长打开会触发外部。线粒体膜破裂、细胞色素c释放和下游细胞激活过去，mPTP 一直是广泛科学研究的中心。几十年来，它仍然是当今生物学中最神秘的现象之一由于其分子组成存在争议且缺乏其孔的结构信息。我们最近证明了 ATP 合酶 c 亚基环在形成然而，mPTP 通道的 ATP 合酶 c 亚基泄漏的门控机制。通道和启动其开放的构象变化尚未被发现。在本提案中，我们将使用单粒子冷冻电子显微镜（cryo-EM）来识别高存在下 ATP 合成酶泄漏通道的解析结构和开放构象我们还将对其中的 ATP 合酶进行原位结构分析。暴露于 Aβ 的初级海马神经元和线粒体内的功能环境在这个项目中，通过使用冷冻电子断层扫描 (cryo-ET) 从 AD 小鼠模型中分离出来。我们还将研究 ATP 合成酶泄漏通道作为新型细胞死亡途径的直接作用在 AD 发病机制中，我们将测试该通道的药理学抑制是否可以挽救该提案的成功完成将揭示 Aβ 诱导的细胞死亡的神经元。线粒体通透性转变的分子机制，ATP 的原子结构合成酶泄漏通道，并将有助于开发针对 ATP 的 AD 新疗法合成。